Visual demonstration of growth hormone receptors on human growth plate chondrocytes.
Visual demonstration of growth hormone receptors on human growth plate chondrocytes.
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人类生长板软骨细胞上生长激素受体的视觉演示。
DOI：
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发表时间：
1990
影响因子：
5.8
通讯作者：
M. J. Waters
中科院分区：
文献类型：
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作者：
G. A. Werther;K. Haynes;Ross Barnard;M. J. Waters
生长激素在人类婴儿体内的作用部位尚不清楚；最近在动物身上的证据表明，生长激素对生长板和其他组织有直接作用。我们使用了一种识别人生长激素受体的单抗来直观地识别和定位人类婴儿生长板中的生长激素受体。取猝死婴儿(n=20)身体胸骨软骨，经脱钙、固定、纵切或胶原酶消化后单层培养软骨细胞。培养的软骨细胞切片用抗生物素-亲和素系统识别人生长激素受体的单抗(MAb 263)进行免疫细胞化学染色。接受胸骨软骨成形术的青少年在手术中也获得了胸骨软骨。在婴幼儿组织中，生长激素受体在增生层和肥厚层的软骨细胞、软骨膜、新生骨中的骨细胞和骨髓中的造血祖细胞的切片中均有表达。培养的软骨细胞生长激素受体呈异质性染色。随着培养时间的延长(5~8d)，染色模式由单个细胞向成组细胞转变。[125I]乳化放射自显影显示人(H)GH与软骨细胞单层有特异性结合(0.6+/-0.3%)。在支持MAb263特异性的情况下，它能够将[125I]hGH从单层置换35%。培养的青少年软骨细胞未能显示出与[125I]hGH的特异性结合。我们得出结论：生长激素受体广泛分布于婴儿生长板中的一系列间充质细胞，包括骨和造血前体细胞。这些受体的表达在完整的组织和培养中似乎都是成熟依赖的，而它们可能在青春期生长高峰期之后不再表达。
The sites of action of GH in the human infant remain unclear; recent evidence in animals suggests direct actions on growth plate and other tissues. We have used a monoclonal antibody recognizing the human GH receptor to visually identify and localize GH receptors in the human infant growth plate. Sternochondral cartilage was obtained at postmortem from infants dying of sudden infant death (n = 20), and either decalcified, fixed, and cut into longitudinal sections or digested with collagenase for monolayer culture of chondrocytes. Sections of cultured chondrocytes were stained immunocytochemically with a monoclonal antibody recognizing human GH receptor (MAb 263), using an avidin-biotin system. Sternochondral cartilage was also obtained at operation from adolescents undergoing sternochondroplasty. In infant tissue, GH receptor was identified in sections in chondrocytes of the proliferative and hypertrophic layers, in perichondrium, in osteocytes in new bone, and in hemopoietic precursor cells in marrow. Cultured chondrocytes showed heterogeneous staining for GH receptor. With prolonged culture from 5-8 days, the pattern of staining changed from individual cells to groups of cells. [125I]Human (h)GH showed specific binding to chondrocyte monolayer (0.6 +/- 0.3%), confirmed visually on emulsion autoradiography. In support of specificity of MAb263, it was able to displace [125I]hGH from monolayers by 35%. Adolescent cultured chondrocytes failed to demonstrate specific binding of [125I]hGH. We conclude that GH receptors are widely distributed in a range of mesenchyme cells in the human infant growth plate, including bone and hemopoietic precursors. The expression of these receptors appears to be maturation dependent in both intact tissue and culture, while they may no longer be expressed after the peak growth phase of puberty.